A major advance in our understanding of acid-base homeostasis and ammonia metabolism is the identification that Rh glycoproteins are ammonia transporters. In the kidney, multiple lines of evidence suggest that Rh glycoprotein C Glycoprotein (Rhcg) is critically important in renal ammonia metabolism. A second advance has been the recognition that Rhcg is expressed in principal cells, a cell not generally known to be involved in acid-base homeostasis, and that principal cell Rhcg expression parallels ammonia excretion. Thus, principal cells may contribute to regulated transcellular ammonia secretion. Finally, Rhcg expression appears to be regulated through post-transcriptional mechanisms. The overall aim of this application is to determine Rhcg's role in acid-base homeostasis and in potassium homeostasis. The first goal is to determine the specific role of Rhcg in the renal response to metabolic acidosis. We will use Cre-loxP technology to generate transgenic mice with kidney-specific, intercalated cell-specific and principal cell-specific Rhcg deletion. We will then examine acid- base homeostasis in these mice under control conditions and in response to metabolic acidosis in order to determine the specific role of Rhcg in renal acid-base homeostasis, and the specific contributions of intercalated cells and principal cells to acid-base homeostasis. Our second aim is to determine Rhcg's specific role in the renal response to hypokalemia. We will again use Cre-loxP technology to generate transgenic mice with kidney- specific, intercalated cell-specific and principal cell-specific Rhcg deletion. We will then examine acid-base and potassium homeostasis in these mice under control conditions and in response to dietary potassium deficiency in order to determine the specific role of Rhcg in the renal response to hypokalemia, and the specific contributions of intercalated cells and principal cells to Rhcg-mediated ion transport in response to hypokalemia. PUBLIC HEALTH RELEVANCE Acid-base disorders are associated with renal stone disease, osteoporosis, muscle atrophy, growth retardation and increased morbidity. The proposed studies will provide new insights into the fundamental mechanisms of acid-base homeostasis, thereby providing underpinnings for new and novel treatments.